- Email: [email protected]
b) polymorphism association with diabetic retinopathy
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Abstracts
Genetics Department, Mashhad University of Medical Science, Mashhad, Iran E-mail addresses: [email protected] (T. Hamzehloei), [email protected] (T.F. Mohajer) Introduction: Thalassemia is an inherited autosomal recessive blood disease that originated in the Mediterranean region. In thalassemia the genetic defect, which could be either mutation or deletion, results in reduced rate of synthesis or no synthesis of one of the globin chains that make up hemoglobin. This can cause the formation of abnormal hemoglobin molecules, thus causing anemia. The basic aim of this study was to identify a suitable molecular approach for prevention of thalassaemia in Mashhad. Material and methods: DNA was extracted from peripheral blood and the whole beta-globin and the entire alpha1 and 2 globin genes were amplified and DNA sequenced. The seven common deletion mutations for apha-globin genes were investigated. Results: Molecular basis of thalassaemia was investigated in 88 mutant alleles. Eighty different mutations were found including one novel allele. Coincidental a-thalassaemia was found in 16% of cases of thalassaemia. Consanguineous marriage and recessive disorders were studied. In 44 couples studied 70% were consanguineous and only 30% were completely unrelated. Discussion: Over 98% of the diagnoses were done by direct mutation analysis. A multiplex polymerase chain reaction for mutation analysis was used and significantly reduced the total cost and time required for prenatal diagnosis. This pilot study appears to have identified a suitable approach for prevention of thalassaemia in Mashhad. Keywords: Thalassemia, Mashhad, Mutation doi:10.1016/j.clinbiochem.2011.08.661
Poster – [A-10-120-1] Association between L55M paraoxonase-1 gene polymorphism and metabolic syndrome Nooshin Sharifia, Tamandani Dor Mohammad Kordia, Mohammad Hashemib, Adam Torkamanzehia a Department of Biology, Faculty of Sciences, University of Sistan and Baluchestan, Zahedan, Iran b Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran E-mail addresses: [email protected] (N. Sharifi), [email protected] (T.D.M. Kordi), [email protected] (M. Hashemi), [email protected] (A. Torkamanzehi) Introduction: Paraoxonase-1 (PON1) is a high-density lipoprotein (HDL) associated enzyme that exhibits antioxidant and anti-atherogenic activities. Three polymorphisms within the PON1 gene affect the enzyme activity. Two of them are located at coding region (L55M, Q192R) and the third one (−108C/T) is placed in promoter region. We looked for a possible link between L55M polymorphism and metabolic syndrome (MES) in samples collected from southeast of Iran. Methods: DNA was extracted from whole blood of 119 patients with MES and 200 healthy controls. The allelic polymorphism at position 55 in the PON1 gene was studied by tetra amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Results: We found the risk of metabolic syndrome in patients with MM and LM + MM genotypes of L55M gene is in marginal border (Odds ratio [OR], 1.33; 95% Confidence Interval [CI], 0.76– 2.31, P = 0.34 and OR, 1.12; 95%CI, 0.68–1.85; P = 0.73 respectively ) with risk of MES.
Conclusion: The present study suggested that the L55M polymorphism is not a major risk factor for MES in the studied sample. Keywords: Paraoxonase-1 (PON1), Metabolic syndrome, Gene, Polymorphism doi:10.1016/j.clinbiochem.2011.08.662
Poster – [A-10-125-1] Nitric oxide synthase 3 VNTR (intron 4 a/b) polymorphism association with diabetic nephropathy Mohseni Mahdieh Mehrab, Bazzaz Javad Tavakkoli, Shirin Hasani-Ranjbar Shariati Hospital, Tehran University, Iran E-mail addresses: [email protected], [email protected] (M.M. Mehrab) Introduction: Nitric oxide synthase3 (NOS3) is involved in several functions playing important role in development of type 2 diabetes mellitus (T2DM), and insulin resistance (IR). It has been demonstrated that NOS3 modulates peripheral and hepatic glucose metabolism and insulin secretion, playing important role in the evolution of IR and T2DM. The aim of this study was to examine the association between NOS3 intron4 VNTR polymorphism and type 2 diabetes in an Iranian population. Patients with diabetes were diagnosed according to American Diabetes Association Criteria. Five CC of peripheral blood was collected in EDTA tubes from patients with type 2 diabetes attending diabetes clinic. Normal healthy controls were from same population. Then DNA was extracted from WBCs, using salting-out method and PCR was performed to determine allele and genotype frequencies for NOS3 gene VNTR polymorphism. A significant difference was found in genotype frequencies of NOS3 polymorphism between patients and controls (aa + ab vs bb p = 0.02, OR = 2.0, 95%CI; 1.05–3.96). Also allele a frequency was significantly increased in patients with diabetes compared with controls (p = 0.007, OR = 2.1, 95%CI; 1.19–4.08). Conclusion: We found a significant difference in distribution of NOS3 polymorphic variants at both allele and genotype frequency level in diabetic patients. A allele is a risk factor of type 2 diabetes in an Iranian population. Keywords: eNOS, Diabetic nephropathy, Polymorphism doi:10.1016/j.clinbiochem.2011.08.663
Poster – [A-10-125-2] Nitric oxide synthase3 VNTR (intron 4 a/b) polymorphism association with diabetic retinopathy Mohseni Mahdieh Mehraba, Shirin Hasani-Ranjbarb, Bazzaz Javad Tavakkolyc a Shariati Hospital, Tehran, Iran b Shariati Hospital, Iran c Department of Medical Genetics, Tehran University of Medical Sciences, Tehran, Iran E-mail addresses: [email protected], [email protected] (M.M. Mehrab) Introduction: Endothelial derived nitric oxide(NO), which is produced by NOS3 participates in several functions related to the vasodilation and mediates vascular action of insulin, and glucose and insulin delivery to the skeletal muscles. High NO levels and endothelial dysfunction have been observed in patients with type 2 diabetes and their first degree relatives as well as in patients with insulin resistance
Abstracts
syndrome. Diabetic retinopathy (DR) is leading cause of blindness in adults between the ages 20 and 65 in industrialized countries. NOS3 haplotype has also shown association with DR in different populations. The aim of this study was to examine the association between NOS3 intron4 VNTR polymorphism and DR in an Iranian population. Diabetic retinopathy was diagnosed by an expert ophthalmologist based on ophthalmoscopic examination. 40 diabetic patients with DR and 96 healthy control subjects were recruited from the same area. PCR was performed to determine allele and genotype frequencies for NOS3 gene VNTR polymorphism. The allele and genotype frequencies of the NOS3 polymorphim conformed to Hardy–Weinberg equilibrium in both patient and control populations. The frequency of a allele was increased in patients with DR compared with healthy controls and also the frequency of genotype aa was increased in patients with DN compared with healthy controls, but there was no significant association between the allele or genotype frequencies of the NO3S polymorphim and patients with diabetic retinopathy. It seems that there is no association between NOS3 gene intron VNTR (4 a/b) polymorphim and DR. Further studies on larger number of samples and on different population are required to confirm the results we observed in this study.
Keywords: NOS3, Diabetic retinopathy, Polymorphism doi:10.1016/j.clinbiochem.2011.08.664
Poster – [A-10-125-3] Study of VPF gene polymorphism association with risk of diabetic retinopathy Mohseni Mahdieh Mehraba, Bazzaz Javad Tavakkolyb, Shirin Hasani-Ranjbara a Endocrinology and Metabolism Research Centre (EMRC), Tehran University of Medical Sciences (TUMS), Shariati Hospital, Tehran, Iran b Department of Medical Genetics, Tehran University of Medical Sciences, Tehran, Iran E-mail addresses: [email protected], [email protected] (M.M. Mehrab) Introduction: VPF (vascular permeability factor) is involved both in regulation of hyperglycemia and structural modifications in diabetes. VPF has a role in accelerating diabetic late complications as diabetic retinopathy (DR) ,nephropathy and neuropathy. In this study we examined VPF gene polymorphism association with DR. Methods: Five CC of peripheral blood was collected in EDTA tubes from patients with type 2 diabetes with and without DR and normal healthy controls. Then DNA was extracted from WBCs, using saltingout method and ARMS-PCR was performed to determine allele and genotype frequencies for VPF −7*C/T and −2578*C/A polymorphism. Results: The frequency of genotype AA was the same in patients with DR compared with diabetic subjects without DR. Also the frequency of A allele was decreased in patients with DR compared to without DFU .There were no significant differences for allele and genotype frequencies of −7*C/T and −2578*C/A polymorphism between diabetic patients with DR and healthy controls. Conclusion: More recently association between this polymorphism and proliferative diabetic retinopathy (PDR) has been found in Japanese population and allele A has been found as a risk factor for development of PDR. Despite detrimental effect of VPF in diabetic retinopathy, it has beneficial effect in diabetic neuropathy. In other studies higher frequency of A allele in patients with DR is conferring a protective effect which might be a result of increased angiogenesis in patients carrying this allele but we were unable to find association between these polymorphisms and DR in Iranian
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population. Maybe more studies on larger population are needed to confirm our results. Keywords: VPF, Diabetic retinopathy, Polymorphism doi:10.1016/j.clinbiochem.2011.08.665
Poster – [A-10-125-4] Evaluation of VPF gene polymorphism association with risk of diabetic nephropathy Mohseni Mahdieh Mehraba, Bazzaz Javad Tavakkolyb a Endocrinology and Metabolism Research Centre (EMRC), Tehran University of Medical Sciences (TUMS), Shariati Hospital, Tehran, Iran b Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran E-mail address: [email protected] (M.M. Mehrab) Introduction: Vascular permeability factor (VPF) is reported to be implicated in the development of diabetic nephropathy (DN). DN is a microvascular complication of diabetes characterized by persistent proteinuria, decreased glomerular filtration rate and increased blood pressure. In this study we performed a candidate gene association study in order to examine VPF gene polymorphism association with DN. Five CC of peripheral blood was collected in EDTA tubes from normal healthy controls and patients with type 2 diabetes with and without DN. Then DNA was extracted from WBCs and ARMS-PCR was performed to determine allele and genotype frequencies for VPF −7*C/T and −2578*C/A polymorphism. The frequency of genotype AA was the same in patients with DN compared with healthy controls (AA vs CA + CC, p = 0.9). The frequency of A allele was increased in patients with DN compared to healthy controls (p = 0.6). There were no significant differences for allele and genotype frequencies of VPF −7*C/T and − 2578*C/A polymorphism between diabetic patients with and without DN. Also there were no significant differences for allele and genotype frequencies of − 7*C/T and − 2578*C/A polymorphism between diabetic patients with DN and healthy controls. We were unable to find association between these polymorphisms and DN in Iranian population. Maybe more studies on larger population are needed to confirm our results. Keywords: VPF, Diabetic nephropathy, Polymorphism
doi:10.1016/j.clinbiochem.2011.08.666
Poster – [A-10-154-1] Differential expression of CXC chemokines CXCL10 (IP-10) and CXCL12 (SDF-1α) in term and pre-term neonates Nabati Saeedeh, Mehdi Mahmoodi, Fariba Aminzadeh, Gholamhossein Hassanshahi Rafsanjan, Iran E-mail addresses: [email protected] (N. Saeedeh), [email protected] (M. Mahmoodi), [email protected] (F. Aminzadeh), [email protected] (G. Hassanshahi) Introduction: Pre-term delivery is a mostly unknown worldwide frequent disorder. The eminent roles of genetic and immunological parameters are assumed to be involved in pre mature delivery. The involvement of chemokine–cytokine network in pre-term labor is documented. This project aimed to investigate the circulating level of CXCL12 (SDF-1 α) and CXCL10 (IP-10) in cord blood of term and preterm delivered fetuses and their mothers.
Abstracts
Genetics Department, Mashhad University of Medical Science, Mashhad, Iran E-mail addresses: [email protected] (T. Hamzehloei), [email protected] (T.F. Mohajer) Introduction: Thalassemia is an inherited autosomal recessive blood disease that originated in the Mediterranean region. In thalassemia the genetic defect, which could be either mutation or deletion, results in reduced rate of synthesis or no synthesis of one of the globin chains that make up hemoglobin. This can cause the formation of abnormal hemoglobin molecules, thus causing anemia. The basic aim of this study was to identify a suitable molecular approach for prevention of thalassaemia in Mashhad. Material and methods: DNA was extracted from peripheral blood and the whole beta-globin and the entire alpha1 and 2 globin genes were amplified and DNA sequenced. The seven common deletion mutations for apha-globin genes were investigated. Results: Molecular basis of thalassaemia was investigated in 88 mutant alleles. Eighty different mutations were found including one novel allele. Coincidental a-thalassaemia was found in 16% of cases of thalassaemia. Consanguineous marriage and recessive disorders were studied. In 44 couples studied 70% were consanguineous and only 30% were completely unrelated. Discussion: Over 98% of the diagnoses were done by direct mutation analysis. A multiplex polymerase chain reaction for mutation analysis was used and significantly reduced the total cost and time required for prenatal diagnosis. This pilot study appears to have identified a suitable approach for prevention of thalassaemia in Mashhad. Keywords: Thalassemia, Mashhad, Mutation doi:10.1016/j.clinbiochem.2011.08.661
Poster – [A-10-120-1] Association between L55M paraoxonase-1 gene polymorphism and metabolic syndrome Nooshin Sharifia, Tamandani Dor Mohammad Kordia, Mohammad Hashemib, Adam Torkamanzehia a Department of Biology, Faculty of Sciences, University of Sistan and Baluchestan, Zahedan, Iran b Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran E-mail addresses: [email protected] (N. Sharifi), [email protected] (T.D.M. Kordi), [email protected] (M. Hashemi), [email protected] (A. Torkamanzehi) Introduction: Paraoxonase-1 (PON1) is a high-density lipoprotein (HDL) associated enzyme that exhibits antioxidant and anti-atherogenic activities. Three polymorphisms within the PON1 gene affect the enzyme activity. Two of them are located at coding region (L55M, Q192R) and the third one (−108C/T) is placed in promoter region. We looked for a possible link between L55M polymorphism and metabolic syndrome (MES) in samples collected from southeast of Iran. Methods: DNA was extracted from whole blood of 119 patients with MES and 200 healthy controls. The allelic polymorphism at position 55 in the PON1 gene was studied by tetra amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Results: We found the risk of metabolic syndrome in patients with MM and LM + MM genotypes of L55M gene is in marginal border (Odds ratio [OR], 1.33; 95% Confidence Interval [CI], 0.76– 2.31, P = 0.34 and OR, 1.12; 95%CI, 0.68–1.85; P = 0.73 respectively ) with risk of MES.
Conclusion: The present study suggested that the L55M polymorphism is not a major risk factor for MES in the studied sample. Keywords: Paraoxonase-1 (PON1), Metabolic syndrome, Gene, Polymorphism doi:10.1016/j.clinbiochem.2011.08.662
Poster – [A-10-125-1] Nitric oxide synthase 3 VNTR (intron 4 a/b) polymorphism association with diabetic nephropathy Mohseni Mahdieh Mehrab, Bazzaz Javad Tavakkoli, Shirin Hasani-Ranjbar Shariati Hospital, Tehran University, Iran E-mail addresses: [email protected], [email protected] (M.M. Mehrab) Introduction: Nitric oxide synthase3 (NOS3) is involved in several functions playing important role in development of type 2 diabetes mellitus (T2DM), and insulin resistance (IR). It has been demonstrated that NOS3 modulates peripheral and hepatic glucose metabolism and insulin secretion, playing important role in the evolution of IR and T2DM. The aim of this study was to examine the association between NOS3 intron4 VNTR polymorphism and type 2 diabetes in an Iranian population. Patients with diabetes were diagnosed according to American Diabetes Association Criteria. Five CC of peripheral blood was collected in EDTA tubes from patients with type 2 diabetes attending diabetes clinic. Normal healthy controls were from same population. Then DNA was extracted from WBCs, using salting-out method and PCR was performed to determine allele and genotype frequencies for NOS3 gene VNTR polymorphism. A significant difference was found in genotype frequencies of NOS3 polymorphism between patients and controls (aa + ab vs bb p = 0.02, OR = 2.0, 95%CI; 1.05–3.96). Also allele a frequency was significantly increased in patients with diabetes compared with controls (p = 0.007, OR = 2.1, 95%CI; 1.19–4.08). Conclusion: We found a significant difference in distribution of NOS3 polymorphic variants at both allele and genotype frequency level in diabetic patients. A allele is a risk factor of type 2 diabetes in an Iranian population. Keywords: eNOS, Diabetic nephropathy, Polymorphism doi:10.1016/j.clinbiochem.2011.08.663
Poster – [A-10-125-2] Nitric oxide synthase3 VNTR (intron 4 a/b) polymorphism association with diabetic retinopathy Mohseni Mahdieh Mehraba, Shirin Hasani-Ranjbarb, Bazzaz Javad Tavakkolyc a Shariati Hospital, Tehran, Iran b Shariati Hospital, Iran c Department of Medical Genetics, Tehran University of Medical Sciences, Tehran, Iran E-mail addresses: [email protected], [email protected] (M.M. Mehrab) Introduction: Endothelial derived nitric oxide(NO), which is produced by NOS3 participates in several functions related to the vasodilation and mediates vascular action of insulin, and glucose and insulin delivery to the skeletal muscles. High NO levels and endothelial dysfunction have been observed in patients with type 2 diabetes and their first degree relatives as well as in patients with insulin resistance
Abstracts
syndrome. Diabetic retinopathy (DR) is leading cause of blindness in adults between the ages 20 and 65 in industrialized countries. NOS3 haplotype has also shown association with DR in different populations. The aim of this study was to examine the association between NOS3 intron4 VNTR polymorphism and DR in an Iranian population. Diabetic retinopathy was diagnosed by an expert ophthalmologist based on ophthalmoscopic examination. 40 diabetic patients with DR and 96 healthy control subjects were recruited from the same area. PCR was performed to determine allele and genotype frequencies for NOS3 gene VNTR polymorphism. The allele and genotype frequencies of the NOS3 polymorphim conformed to Hardy–Weinberg equilibrium in both patient and control populations. The frequency of a allele was increased in patients with DR compared with healthy controls and also the frequency of genotype aa was increased in patients with DN compared with healthy controls, but there was no significant association between the allele or genotype frequencies of the NO3S polymorphim and patients with diabetic retinopathy. It seems that there is no association between NOS3 gene intron VNTR (4 a/b) polymorphim and DR. Further studies on larger number of samples and on different population are required to confirm the results we observed in this study.
Keywords: NOS3, Diabetic retinopathy, Polymorphism doi:10.1016/j.clinbiochem.2011.08.664
Poster – [A-10-125-3] Study of VPF gene polymorphism association with risk of diabetic retinopathy Mohseni Mahdieh Mehraba, Bazzaz Javad Tavakkolyb, Shirin Hasani-Ranjbara a Endocrinology and Metabolism Research Centre (EMRC), Tehran University of Medical Sciences (TUMS), Shariati Hospital, Tehran, Iran b Department of Medical Genetics, Tehran University of Medical Sciences, Tehran, Iran E-mail addresses: [email protected], [email protected] (M.M. Mehrab) Introduction: VPF (vascular permeability factor) is involved both in regulation of hyperglycemia and structural modifications in diabetes. VPF has a role in accelerating diabetic late complications as diabetic retinopathy (DR) ,nephropathy and neuropathy. In this study we examined VPF gene polymorphism association with DR. Methods: Five CC of peripheral blood was collected in EDTA tubes from patients with type 2 diabetes with and without DR and normal healthy controls. Then DNA was extracted from WBCs, using saltingout method and ARMS-PCR was performed to determine allele and genotype frequencies for VPF −7*C/T and −2578*C/A polymorphism. Results: The frequency of genotype AA was the same in patients with DR compared with diabetic subjects without DR. Also the frequency of A allele was decreased in patients with DR compared to without DFU .There were no significant differences for allele and genotype frequencies of −7*C/T and −2578*C/A polymorphism between diabetic patients with DR and healthy controls. Conclusion: More recently association between this polymorphism and proliferative diabetic retinopathy (PDR) has been found in Japanese population and allele A has been found as a risk factor for development of PDR. Despite detrimental effect of VPF in diabetic retinopathy, it has beneficial effect in diabetic neuropathy. In other studies higher frequency of A allele in patients with DR is conferring a protective effect which might be a result of increased angiogenesis in patients carrying this allele but we were unable to find association between these polymorphisms and DR in Iranian
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population. Maybe more studies on larger population are needed to confirm our results. Keywords: VPF, Diabetic retinopathy, Polymorphism doi:10.1016/j.clinbiochem.2011.08.665
Poster – [A-10-125-4] Evaluation of VPF gene polymorphism association with risk of diabetic nephropathy Mohseni Mahdieh Mehraba, Bazzaz Javad Tavakkolyb a Endocrinology and Metabolism Research Centre (EMRC), Tehran University of Medical Sciences (TUMS), Shariati Hospital, Tehran, Iran b Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran E-mail address: [email protected] (M.M. Mehrab) Introduction: Vascular permeability factor (VPF) is reported to be implicated in the development of diabetic nephropathy (DN). DN is a microvascular complication of diabetes characterized by persistent proteinuria, decreased glomerular filtration rate and increased blood pressure. In this study we performed a candidate gene association study in order to examine VPF gene polymorphism association with DN. Five CC of peripheral blood was collected in EDTA tubes from normal healthy controls and patients with type 2 diabetes with and without DN. Then DNA was extracted from WBCs and ARMS-PCR was performed to determine allele and genotype frequencies for VPF −7*C/T and −2578*C/A polymorphism. The frequency of genotype AA was the same in patients with DN compared with healthy controls (AA vs CA + CC, p = 0.9). The frequency of A allele was increased in patients with DN compared to healthy controls (p = 0.6). There were no significant differences for allele and genotype frequencies of VPF −7*C/T and − 2578*C/A polymorphism between diabetic patients with and without DN. Also there were no significant differences for allele and genotype frequencies of − 7*C/T and − 2578*C/A polymorphism between diabetic patients with DN and healthy controls. We were unable to find association between these polymorphisms and DN in Iranian population. Maybe more studies on larger population are needed to confirm our results. Keywords: VPF, Diabetic nephropathy, Polymorphism
doi:10.1016/j.clinbiochem.2011.08.666
Poster – [A-10-154-1] Differential expression of CXC chemokines CXCL10 (IP-10) and CXCL12 (SDF-1α) in term and pre-term neonates Nabati Saeedeh, Mehdi Mahmoodi, Fariba Aminzadeh, Gholamhossein Hassanshahi Rafsanjan, Iran E-mail addresses: [email protected] (N. Saeedeh), [email protected] (M. Mahmoodi), [email protected] (F. Aminzadeh), [email protected] (G. Hassanshahi) Introduction: Pre-term delivery is a mostly unknown worldwide frequent disorder. The eminent roles of genetic and immunological parameters are assumed to be involved in pre mature delivery. The involvement of chemokine–cytokine network in pre-term labor is documented. This project aimed to investigate the circulating level of CXCL12 (SDF-1 α) and CXCL10 (IP-10) in cord blood of term and preterm delivered fetuses and their mothers.